• Biomedical Science Letters
• /
• v.20 no.4
• /
• pp.244-249
• /
• 2014

Arazyme is a metalloprotease secreted by Aranicola proteolyticus that was previously shown to suppress cytokine expression of keratinocytes and endothelial cells and inhibit histopathological features in an atopic dermatitis-like animal model. However, the regulatory effects of arazyme in other allergic diseases have yet to be elucidated. In this study, we investigated whether arazyme is effective against neutrophil apoptosis in allergic diseases such as allergic rhinitis and asthma. Arazyme inhibited neutrophil apoptosis of normal subjects in a dose-dependent manner. However, the antiapoptotic effect of arazyme was reversed by LY294002, an inhibitor of PI3K, AKTi, an inhibitor of Akt, PD98059, an inhibitor of MEK, and BAY-11-7085, an inhibitor of NF-${\kappa}B$. Arazyme induced activation of NF-${\kappa}B$ via PI3K/Akt/ERK pathway. The anti-apoptotic effect of arazyme is associated with inhibition of cleavage of caspase 3 and caspase 9. Arazyme inhibited constitutive apoptosis of neutrophil in a dose-dependent manner in allergic subjects, and its mechanism was shown to be associated with PI3K/Akt/ERK/NF-${\kappa}B$. The results presented here improve our understanding of neutrophil apoptosis regulation and will facilitate development of drugs for treatment of allergic diseases.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.48 no.2
• /
• pp.67-77
• /
• 2024

This study aims to assess the efficacies of exogenous enzymes, derived from invertebrate gut-associated microbes, as feed additives, in reducing volatile organic compound (VOC) emissions using an in vitro pig intestine continuous fermentation system. An in vitro continuous fermentation model was used to simulate a comparable bionic digestion system by co-reacting feed, enzymatic additives (arazyme, mannanase, and xylanase, derived from the gut bacteria of Nephila clavata, Eisenia fetida, and Moechotypa diphysis, respectively), and gastrointestinal microbes, followed by an analysis of their correlations. A significant correlation was observed between exogenous enzyme supplementation and reduced VOC emissions in the fecal phase of continuous fermentation (p < 0.05). The concentration of VOCs decreased by 3.75 and 2.75 ppm in the treatment group following arazyme and multi-enzyme supplementation, respectively, compared to that in the control group (7.83 ppm). In addition, supplementation with arazyme and multiple enzymes significantly affected the microbial composition of each fermentation phase (p < 0.05). In particular, Lactiplantibacillus pentosus and Pediococcus pentosaceus, which changed in abundance according to arazyme or multi-enzyme supplementation, exhibited a positive relationship with VOC emissions. These results suggest that exogenous enzymes derived from invertebrate gut-associated bacteria can be efficiently applied as feed additives, leading to a reduction in VOC emissions.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.5
• /
• pp.761-768
• /
• 2007

Serratia proteamaculans HY-3 isolated from the digestive tract of a spider produces an extracellular protease named arazyme, with an estimated molecular mass of 51.5 kDa. The purified enzyme was characterized as having high activities at wide pH and temperature ranges. We further characterized biochemical features of the enzymatic reactions under various reaction conditions. The protease efficiently hydrolyzed a broad range of protein substrates including albumin, keratin, and collagen. The dependence of enzymatic activities on the presence of metal ions such as calcium and zinc indicated that the enzyme is a metalloprotease, together with the previous observation that the proteolytic activity of the enzyme was not inhibited by aspartate, cysteine, or serine protease inhibitors, but strongly inhibited by 1,10-phenanthroline and EDTA. The araA gene encoding the exoprotease was isolated as a 5.6 kb BamHI fragment after PCR amplification using degenerate primers and subsequent Southern hybridization. The nucleotide sequence revealed that the deduced amino acid sequences shared extensive similarity with those of the serralysin family of metalloproteases from other enteric bacteria. A gene(inh) encoding a putative protease inhibitor was also identified immediately adjacent to the araA structural gene.

• Korean Journal of Poultry Science
• /
• v.36 no.1
• /
• pp.47-55
• /
• 2009

This study was carried out to investigate the effects of spider-derived protease [Arazyme (AZ)] supplementation on egg production, egg quality, cecal microflora and viscosity of intestinal contents in laying hens at the late production. A total of two hundred, 59-wks-old, Hy-Line Brown layers were divided into four groups, placed into 5 replicates per group (50 birds per group) and fed one of four diets containing 0.05, 0.075 or 0.1% AZ or control for 5 wks. The results showed that egg weight in group fed the diet containing 0.05% AZ was significantly higher than those of other groups (P<0.05). There were no significant differences in egg interior and eggshell qualities among the groups. The serum parameters such as total cholesterol, the levels of GOT and GPT were not influenced by AZ feeding. There were no significant differences in the cecal number of total microbes, Coliforms and lactic acid bacteria among the groups. The viscosity and ammonia concentration of intestinal contents in the groups fed AZ were significantly reduced compared to those of the control (P<0.05). In conclusion, laying hens fed AZ supplemented diet improved egg weight as compared with that of control. In addition, the AZ supplemented diet reduced the viscosity and ammonia concentration of intestinal contents, reflecting a significant feed enzyme effect.